Conformational Dynamics, Intramolecular Domain Conformation Signaling, and Activation of Apo-FimD Revealed by Single-Molecule Fluorescence Resonance Energy Transfer Studies

doi:10.1021/acs.biochem.9b00080

	Conformational Dynamics, Intramolecular Domain Conformation Signaling, and Activation of Apo-FimD Revealed by Single-Molecule Fluorescence Resonance Energy Transfer Studies
	Liu, Yanqing 1,2; Sun, Chuanqi 1,2; Han, Long 1,2; Yu, Yuqi 3; Zhou, Haizhen 2; Shao, Qiang 3; Lou, Jizhong 1,4; Zhao, Yongfang 1,2; Huang, Yihua 1,2
刊名	BIOCHEMISTRY
	2019-04-09
卷号	58 期号:14 页码:1931-1941
ISSN号	0006-2960
DOI	10.1021/acs.biochem.9b00080
通讯作者	Huang, Yihua(yihuahuang@sun5.ibp.ac.cn)
英文摘要	The chaperone-usher secretion pathway is a conserved bacterial protein secretion system dedicated to the biogenesis of adhesive fibers. Usher, a multidomain-containing outer membrane protein, plays a central role in this process by acting as a molecular machine that recruits different chaperone- subunit complexes, catalyzes subunit polymerization, and forms a channel for secretion of the assembled subunits. While recent crystal structural studies have greatly advanced our understanding of the structure and function of ushers, the overall architecture of the full-length apo-usher, the molecular events that dictate conformational changes in usher during pilus biogenesis, and its activation by the specific chaperone-adhesin complex remain largely elusive. Using single-molecule fluorescence resonance energy transfer studies, we found that the substrate-free usher FimD (apo-FimD) adopts a contracted conformation that is distinct from its substrate-bound states; both the N-terminal domain (NTD) and the C-terminal domain (CTD) of apo-FimD are highly dynamic, and FimD coordinates its domain conformational changes via intramolecular domain conformation signaling. By combining these studies with in vitro photo-cross-linking studies, we further show that only the chaperone-bound adhesin (FimC:FimH) can be transferred to the CTD, dislocates the plug domain, and triggers conformational changes in the remaining FimD domains. Taken together, these studies delineate an overall architecture of the full-length apo-FimD, provide detailed mechanic insight into the activation of apo-FimD, and explain why FimD could adjust its conformational states to perform multiple functions in each cycle of pilus subunit addition and ensure that pilus assembly proceeds progressively in a cellular energy-free environment.
资助项目	Ministry of Science and Technology[2016YFA0500404] ; National Natural Science Foundation of China[31625009] ; National Natural Science Foundation of China[31470743] ; Strategic Priority Research Program of the Chinese Academy of Sciences[XDB080203]
WOS关键词	CHAPERONE-SUBUNIT COMPLEXES ; BACTERIAL OUTER-MEMBRANE ; DONOR-STRAND EXCHANGE ; TYPE-1 PILI ; CRYSTAL-STRUCTURE ; STRUCTURAL BASIS ; PAPC USHER ; BIOGENESIS ; ADHESIN ; RECOGNITION
WOS研究方向	Biochemistry & Molecular Biology
语种	英语
出版者	AMER CHEMICAL SOC
WOS记录号	WOS:000464476500013
内容类型	期刊论文
源URL	[http://119.78.100.183/handle/2S10ELR8/290129]
专题	中国科学院上海药物研究所
通讯作者	Huang, Yihua
作者单位	1.Univ Chinese Acad Sci, Beijing 100101, Peoples R China 2.Chinese Acad Sci, Inst Biophys, CAS Ctr Excellence Biomacromol, Natl Lab Biomacromol, 15 Datun Rd, Beijing 100101, Peoples R China 3.Chinese Acad Sci, Shanghai Inst Mat Med, CAS Key Lab Receptor Res, Drug Discovery & Design Ctr, 555 Zuchongzhi Rd, Shanghai 201203, Peoples R China 4.Chinese Acad Sci, Inst Biophys, Key Lab RNA Biol, Beijing 100101, Peoples R China
推荐引用方式 GB/T 7714	Liu, Yanqing,Sun, Chuanqi,Han, Long,et al. Conformational Dynamics, Intramolecular Domain Conformation Signaling, and Activation of Apo-FimD Revealed by Single-Molecule Fluorescence Resonance Energy Transfer Studies[J]. BIOCHEMISTRY,2019,58(14):1931-1941.
APA	Liu, Yanqing.,Sun, Chuanqi.,Han, Long.,Yu, Yuqi.,Zhou, Haizhen.,...&Huang, Yihua.(2019).Conformational Dynamics, Intramolecular Domain Conformation Signaling, and Activation of Apo-FimD Revealed by Single-Molecule Fluorescence Resonance Energy Transfer Studies.BIOCHEMISTRY,58(14),1931-1941.
MLA	Liu, Yanqing,et al."Conformational Dynamics, Intramolecular Domain Conformation Signaling, and Activation of Apo-FimD Revealed by Single-Molecule Fluorescence Resonance Energy Transfer Studies".BIOCHEMISTRY 58.14(2019):1931-1941.